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A blood preservation agent for protecting free dna and its application

A technology of preservatives and blood, applied in the field of blood preservatives for the protection of free DNA, can solve the problems of harm to the human body, etc., achieve the effects of preventing pollution, reducing storage conditions, and saving economic costs

Active Publication Date: 2018-07-31
广州奇辉生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patent application number 201510132774.4 discloses a formaldehyde release agent in the blood anticoagulant component used to protect free DNA. The formaldehyde produced by the formaldehyde is a toxic substance and will be harmful to the human body.

Method used

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  • A blood preservation agent for protecting free dna and its application
  • A blood preservation agent for protecting free dna and its application
  • A blood preservation agent for protecting free dna and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1 room temperature static preservation experiment

[0061] The blood preservation agent 1 provided in this embodiment, every 100 milliliters of the protective agent 1 includes the following component weight (g):

[0062] Dipotassium EDTA

8.8g

ε-Polylysine

1.8g

urea

8.0g

glucose

9.6g

Mannitol

5.0g

adenine

0.3g

[0063] Preparation method: prepare 100ml of blood preservative 1, calculate according to the concentration of the above components, and prepare materials: dipotassium edetate: 8.8g, ε-polylysine: 1.8g, urea: 8.0g, glucose: 9.6g , Mannitol: 5.0g, Adenine: 0.3g.

[0064] The above mixture was fixed to 100ml, 0.05mol / l, pH7.4 tris-hydrochloric acid buffer solution, and finally the finished blood preservation agent 1 was obtained.

[0065] When the blood preservative 1 obtained in this embodiment is used as a sample free DNA preservative when collecting blood samples, transporting blood s...

Embodiment 2

[0104] Example 2 Transport experiment at normal temperature:

[0105] The components and ratios of blood preservation agent 1 and blood preservation agent 2 provided in this embodiment are the same as those in embodiment 1.

[0106] (1) Experimental method:

[0107] Vacuum blood collection tubes (1 containing 125 μl preservative 1) were used to draw 2 tubes of 5ml blood samples from each of the 6 volunteers, and mixed upside down 10 times to make the blood and the preservative into a uniform system. The blood samples of each volunteer were collected for 0 days (within 6 hours) and transported at room temperature for 4 days (96 hours), respectively. One tube was taken for plasma separation, and only 600 μl of plasma was taken for each tube as a plasma sample. The method of plasma separation Can be with embodiment 1.

[0108] Extraction of plasma cell-free DNA: Extract plasma cell-free DNA from 600 μl plasma samples of each volunteer (the extraction kit can be selected from th...

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Abstract

The invention belongs to preservation agents of blood samples and in particular relates to a blood preservation agent for protecting free DNA. Every 100ml of the preservation agent comprises the following components: 6.0-8.8g of an anticoagulant, 0.1-2.0g of a natural preservative, 4.8-11.7g of a DNA enzyme inhibitor, 5.0-10.0g of glucose, 5.0-8.0g of an antihemolytic agent, 0.1-0.5g of purine and the balance of buffer solution, totaling 100ml. The preservation agent disclosed by the invention can achieve the effects of effectively preventing blood cells from rupturing to release genome DNA so as to cause pollution, preventing degradation of free DNA in the plasma and providing a stable environment for the free DNA in the plasma. Moreover, the preservation agent is safe and non-toxic to the human body and environment and is favorable for accurate detection and analysis of the free DNA.

Description

technical field [0001] The invention belongs to a blood sample preservation reagent, in particular to a blood preservation reagent for protecting free DNA and an application thereof. Background technique [0002] At present, there are more and more clinical applications of plasma free DNA. For example, non-invasive prenatal genetic testing and tumor diagnosis both use plasma free DNA as the detection target. During the detection process, the amount of plasma free DNA is an important detection index. The content of free DNA in human plasma is very small, which is easily degraded by DNase in the blood, and is easily polluted by genomic DNA released by leukocyte lysis, which seriously affects the test results. [0003] In order to solve these problems, the main measures taken at this stage are: (1) Select blood collection tubes with anticoagulants (EDTA, heparin, citrate) to collect blood samples to prevent coagulation of whole blood cells and reduce the release of genomic DNA...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/10
Inventor 朱奇刘伟华胡文晖王灵敏李岩松
Owner 广州奇辉生物科技有限公司
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